Generating Yeast Two-Hybrid Bait Strains
Program in Systems Biology; Program in Molecular Medicine
Biochemistry, Biophysics, and Structural Biology | Genetics and Genomics | Laboratory and Basic Science Research | Systems Biology
Generating DNA-binding domain (DB)-bait strains for Gateway-compatible yeast two-hybrid (Y2H) screens involves three steps. The first is to generate an Entry clone containing a DNA fragment encoding the protein of interest (e.g., an open reading frame, ORF). The second is to transfer this DNA fragment from the Entry clone to the Y2H Destination vector, pDEST32. The final step is to transform this construct into the Y2H yeast strain, MaV103. This protocol takes 24-37 d plus sequence confirmation, if necessary, to complete.
DOI of Published Version
Cold Spring Harb Protoc. 2018 Jul 2;2018(7):pdb.prot094979. doi: 10.1101/pdb.prot094979. Link to article on publisher's site
Cold Spring Harbor protocols
Reece-Hoyes, John S. and Walhout, Albertha J. M., "Generating Yeast Two-Hybrid Bait Strains" (2018). Program in Systems Biology Publications and Presentations. 143.